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1.
The occurrence and geographic distribution of longidorid nematode species inhabiting the rhizosphere of cultivated olive (cvs. Chemlali and Chétoui) in Tunisia were investigated. Morphological and morphometrical studies identified three Longidorus and six Xiphinema species, with frequencies of prevalence as following: Longidorus africanus (23.0 %), L. euonymus (4.5 %), L. glycines (13.7 %), Xiphinema conurum (13.7 %), X. italiae (36.4 %), X. meridianum (13.7 %), X. pachtaicum (18.2 %), X. robbinsi (9.1 %), and Xiphinema sp. (4.5 %). The three Longidorus species were reported for the first time in Tunisia, in addition to two species of Xiphinema (viz. X. meridianum and X. robbinsi). Molecular characterisation using D2-D3 expansion regions of 28S rRNA and ITS1-rRNA was carried out and Bayesian inference analysis was used to reconstruct phylogenetic relationships among these species and with other longidorids. Twenty-five new D2-D3 of 28S rRNA gene sequences were obtained in the present study, seven for Longidorus and 18 for Xiphinema spp., as well as 14 new ITS1 rRNA gene sequences (seven for Longidorus and seven for Xiphinema spp.).  相似文献   

2.
Populations of Xiphinema brevicollum occurring in the Czech Republic were described morphologically and molecularly. Published species-specific primer set BL18 and BV3 was used to amplify three populations of X. brevicollum from the Czech Republic. These primers were tested against 9 species of Xiphinema and 11 species of Longidorus. Amplification was also observed for X. inaequale, X. italiae and X. lambertii. Three additional markers, mitochondrial cytochrome c oxidase subunit 1, ribosomal D2/D3 expansion segment of the 28S gene and 18S gene, were amplified and sequenced for X. brevicollum, X. inaequale and X. lambertii belonging to X. americanum-group. Comparison of cox1 sequences of X. brevicollum from the Czech Republic with X. taylori from the Slovak Republic (accession number AM086702) suggested that these populations represent the same species.  相似文献   

3.
The occurrence and geographic distribution of longidorid nematode species inhabiting the rhizosphere of cultivated and wild olive and grapevine in Crete Island were investigated. Morphological and morphometrical studies identified five Longidorus and six Xiphinema species, with frequencies of prevalence (for wild and cultivated olives and grapevines, respectively) as follows: Longidorus closelongatus (2.0–13.3 %), L. cretensis (1.0–6.7 %), L. moesicus (13.3 % only in grapevines), L. orientalis (3.3 % only in grapevines), L. pseudoelongatus (7.0 % only in olives), Xiphinema cretense n. sp. (3.0 % only in olives), X. index (3.0–23.3 %), X. israeliae (6.3 % only in olives), X. italiae (3.3–10.0 %), X. pachtaicum (26.7–42 %) and X. simile (3.3 % only in grapevines). Xiphinema cretense n. sp. is characterized by a body size 3,872–6,135 μm long, lip region anteriorly rounded, separated from the rest of the body by a depression, odontostyle and odontophore 140.6 and 80.3 μm long respectively, vulva position at 46.0–50.5 %, female tail 31.0–38.0 μm long, nearly hemispherical with curvature essentially dorsal and with a tip completely rounded or presenting a very short bulge, c ratio (119.1–186.9), c’ ratio (0.7–0.8). Molecular characterisation using D2-D3 expansion regions of 28S rRNA, 18S rRNA and ITS1-rRNA was carried out and maximum likelihood and Bayesian inference analysis were used to reconstruct phylogenetic relationships among these species and with other longidorids.  相似文献   

4.
The occurrence and geographic distribution of longidorid nematode species inhabiting the rhizosphere of grapevine plants in southern Spain were investigated. Nematode surveys were conducted on 77 vineyards during the spring seasons of 2006, 2007 and 2008 in the main Andalusian grapevine-growing areas, including the provinces of Cádiz, Córdoba, and Huelva. Morphological and morphometrical studies identified two Longidorus and nine Xiphinema species, viz.: Longidorus alvegus, L. magnus, Xiphinema adenohystherum, X. hispidum, X. index, X. italiae, X. lupini, X. nuragicum, X. pachtaicum, X. rivesi, and X. turcicum. Overall, frequencies of infestation were, in decreasing order: X. pachtaicum 90.8%, X. index 30.3%, X. italiae 13.2%, L. magnus 11.8%, X. hispidum 7.9%, X. lupini 3.9%, L. alvegus and X. rivesi 2.6%, and X. adenohystherum, X. nuragicum and X. turcicum 1.3%. Xiphinema hispidum, X. lupini, L. alvegus and L. magnus were compared with nematode type specimens and are reported for the first time in Spain. Furthermore, the male of L. alvegus is described for the first time in the literature. Molecular characterisation of these species using D2–D3 expansion regions of 28S rRNA, 18S rRNA and ITS1-rRNA was carried out and maximum likelihood and Bayesian inference analysis were used to reconstruct phylogenetic relationships among these species and with other longidorids. The monophily of the genera Xiphinema and Longidorus was accepted and the genera Paralongidorus and Xiphidorus were rejected by the Shimodaira-Hasegawa test based on tree topologies.  相似文献   

5.
The ectoparasitic dagger nematodes Xiphinema index and Xiphinema diversicaudatum, often at low numbers in the soil, are vectors of grapevine nepoviruses, which cause huge agronomical problems for the vineyard industry. This study reports a method, based on real‐time PCR, for the specific detection of these species and of the closely related non‐vector species Xiphinema vuittenezi and Xiphinema italiae. Specific primers and TaqMan probes were designed from the ribosomal DNA internal transcribed spacer 1 (ITS1), enabling the specific detection of single individuals of each of the X. index, X. diversicaudatum, X. italiae and X. vuittenezi species whatever the nematode population. The specificity of detection and absence of false positive reaction were confirmed in samples of each species mixed with the three other Xiphinema species or mixed with nematodes representative from other genera (non‐plant‐parasitic Dorylaimida, Longidorus sp., Meloidogyne spp., Globodera spp. and Pratylenchus sp.). The method was shown to be valid for the relative quantification of X. index numbers through its use, from crude nematode extracts of soil samples, in a greenhouse assay of grapevine accessions ranging from highly susceptible to resistant. As an alternative to time‐consuming microscopic identification and counting, this real‐time PCR method will provide a fast, sensitive and reliable diagnostic and relative quantification technique for X. index nematodes extracted from fields or controlled conditions.  相似文献   

6.
Principal components and hierarchical cluster analysis based on morphometric characters of the type populations of the 39 species attributed to the Xiphinema americanum-group confirmed the occurrence of five subgroups. The use of a single character is generally insufficient to characterize a species because of the wide intra-population and intra-specific variations. Hierarchical cluster analysis, although impractical for species/population identification, appears useful in the study of the relationships between species or groups of species.  相似文献   

7.
A new dagger nematode, Xiphinema tica n. sp., is described and illustrated from several populations extracted from soil associated with several crops and wild plants in Costa Rica. The new dagger nematode is characterised by a moderate body size (3276–4240 μm), a rounded lip region, ca 13.5 μm wide, separated from body contour by a shallow depression, amphidial fovea large, stirrup-shaped, a moderately long odontostyle ca 135 μm long, stylet guiding ring located at ca 122 μm from anterior end, vulva almost equatorial (50–54%), well-developed Z-organ, with heavy muscularised wall containing in the most of specimens observed two moderately refractive inclusions variable in shape (from round to star-shaped), with uterine spines and crystalloid bodies; female tail short, dorsally convex-conoid, with rounded end and a small peg, with a c’ ratio ca 0.8, bearing two or three pairs of caudal pores and male absent. The unique and novel uterine differentiation based on the coexistence of a well-developed Z-organ mixed with uterine spines and crystalloid bodies in Xiphinema prompted us to update and include this combination of characters in the polytomous key of Loof and Luc (1990). Integrative diagnosis was completed with molecular data obtained, using D2-D3 expansion segments of 28S rDNA, ITS1-rDNA, partial 18S–rDNA and the partial mitochondrial gene cytochrome c oxidase subunit 1 (coxI). The phylogenetic relationships of this species with other Xiphinema spp. indicated that X. tica n. sp. was monophyletic to the other species from the morphospecies Group 4, Xiphinema oleae.  相似文献   

8.
Gut bacteria of fruit fly, Bactrocera tau (Walker) (Diptera: Tephritidae), were isolated and the isolates attractive to B. tau adults were characterized using morphological, biochemical and 16S rRNA analyses to determine their taxonomic position. Based upon morphological, biochemical and 16S rRNA sequences (on the basis of closest match), five gut bacterial species of B. tau were characterized as Delftia acidovorans, Pseudomonas putida, Flavobacterium sp., Defluvibacter sp. and Ochrobactrum sp., of which four bacterial isolates, viz., Delftia acidovorans, Flavobacterium sp., Defluvibacter sp. and Ochrobactrum sp. are new records from guts of the fruit fly species.  相似文献   

9.
A new mono-sexual root-lesion nematode species, Pratylenchus oleae n. sp., parasitizing roots of olive plants cv. Koroneiki in commercial fields at Ouled Chamekh (central Tunisia), and wild and cultivated olive (cv. Picual) plants in Agua Amarga (southern Spain) is described. The new species is characterised by the female having a lip region slightly offset and bearing three annuli, stylet 16.5 (14.5-17.0) μm long, with prominent rounded knobs, pharyngeal overlapping rather long (22–36) μm, lateral fields areolated and with four incisures and diagonal lines in middle band, spermatheca rounded but non-functional, tail short, conoid-rounded to subcylindrical, usually annulated terminus, males unknown, and a specific D2-D3, ITS1, 18S-rRNA, hsp90 and COI sequences. Morphologically this species is related to P. cruciferus, P. delattrei, and P. kumamotoensis. The results of the phylogenetic analysis based on sequences of the D2-D3 expansion regions of 28S, partial 18S and ITS rRNA genes confirmed the close relationship of P. oleae n. sp. with P. dunensis, P. penetrans, P. pinguicaudatus, from which was clearly separated. A PCR-based diagnostic assay was also developed for identification of P. oleae n. sp. using the species-specific primers Poleae_fw1_4 and Poleae_rv1 that amplify a 547-bp fragment in the internal transcribed spacer (ITS1) region of ribosomal DNA, which clearly separate from other root-lesion nematodes damaging olive such as P. penetrans and P. vulnus.  相似文献   

10.
The needle nematodes of the genus Longidorus can cause diseases of various crops and trees, and are comprised of more than 150 valid species. Eleven valid and six unidentified species of the genus Longidorus collected in different regions of Russia, two states of USA, Germany, New Zealand and Ukraine were molecularly characterized using analysis of the partial 18S rRNA and the D2–D3 expansion segments of the 28S rRNA gene sequences. Fifty-four partial 28S rRNA and fifteen partial 18S rRNA gene sequences were obtained for the present study. Using molecular criteria, we confirmed the morphological identification and distinguished between the following species: L. aetnaeus, L. africanus, L. andalusicus, L. artemisiae, L. caespiticola, L. distinctus, L. elongatus, L. euonymus, L. intermedius, L. leptocephalus and L. lignosus. Two longidorid populations from Russia and four from California were not identified to a species level. We obtained the full length D2–D3 of 28S rRNA gene sequence from several freshly-collected L. artemisiae samples. We confirmed the identity of the D2 region of 28S rRNA gene sequence with a short D2 of 28S rRNA gene fragment sequence previously obtained from formalin-fixed nematodes embedded in the L. artemisiae paratype slides. Longidorus lignosus was molecularly characterized and L. aetnaeus was reported from Russia for the first time. PCR-D2-D3-RFLP diagnostic profiles generated by five restriction enzymes: AluI, HinfI, Bsp143I, Tru1I and RsaI are presented for sixteen Longidorus species.  相似文献   

11.
Meloidogyne incognita is one of the most polyphagous species of root-knot nematodes occurring in Brazil and worldwide. Eight M. incognita isolates were studied, representing two enzymatic phenotypes (esterase and malate desydrogenase: I1/N1, I2/N1) and four cryptic Meloidogyne sp.1 (S2/N1) isolates, representing one cytological type (3n?=?40–46). Three M. hispanica isolates (Hi3/N1, 2n?=?32–36) and two of an atypical Meloidogyne sp.2 (S2a/N3, 3n?=?40–44) were included in this study for comparison. All isolates were tested with three M. incognita-specific molecular markers. The primer pairs B06F/R, miF/R and incK14F/R amplified three species-specific fragments of 1,200?bp, 955?bp and 399?bp, respectively for M. incognita and Meloidogyne sp.1 isolates. No amplification occurred in the M. hispanica and Meloidogyne sp.2 isolates, except with primers miF/R (1,650?bp). The genetic variability of the Meloidogyne spp. isolates was evaluated, using RAPD and ISSR markers. The phylogenetic analyses revealed two strongly supported monophyletic clades: clade I, consisting of M. hispanica and the atypical Meloidogyne sp.2 isolates, and clade II, clustering together all M. incognita and the Meloidogyne sp.1 isolates. Considering the biometrical, cytological and molecular approaches, it was possible to conclude that the isolates with three enzymatic phenotypes (I1/N1, I2/N1 and S2/N1) presented the characteristics described for M. incognita. Some correlations were detected between the isozymatic phenotypes and the tree topology (S2a/N3, Hi3/N1, I1/N1, S2/N1), but no strict correlation could be observed for the phenotype I2/N1 and one isolate of S2/N1. Morphologically, the Msp.2 isolates differ from M. incognita and M. hispanica by the female stylet features presenting straight cone tip and round pear shaped knobs, posteriorly sloping. The results of this study suggested that the Msp.2 isolates with phenotypes S2aN3 belong to a new or an unidentified species closely related to M. hispanica.  相似文献   

12.
Ocimum (Lamiaceae) is an important plant genus, with many species used for food flavorings and for essential oils. Fusarium wilt (Fusarium oxysporum f. sp. basilici, FOB) is the most important disease of basil (O. basilicum L.). Twenty-five accessions of O. basilicum, five of O. americanum and two of O. campechianum were initially evaluated for resistance to one FOB isolate (named as FOB-1). Eight accessions (identified as resistant to FOB-1) and one susceptible control were reevaluated to FOB-1 and four other FOB isolates of distinct geographic origins. The FOB isolates varied in aggressiveness and interacted differentially with the Ocimum accessions. Two accessions of O. americanum, one of O. campechianum, and one of O. basilicum had high levels of resistance to all five FOB isolates. The Ocimum germplasm identified here could represent useful sources of resistance genes for developing cultivars with wide-spectrum resistance (i.e., effective against a broad range of FOB isolates). In addition, having a set of potential differential accessions might be useful for large-scale analysis of FOB isolates to demonstrate the presence of physiological races in the Ocimum–FOB pathosystem.  相似文献   

13.
Regions within the mitochondrial gene encoding for the nicotinamide dehydrogenase subunit 4 (nad4) were characterized to evaluate the extent of genetic variation within and among Xiphinema diversicaudatum and Xiphinema simile populations. Four different sequence variants of nad4 were determined among eight populations of X. diversicaudatum and three variants among three populations of X. simile. Nucleotide variation was detected in 28 of 411 bp (2.43 to 4.87 %) in X. diversicaudatum and in three of 395 bp (0.25 to 0.76 %) in X. simile. This represents the first study based on the characterization of the nad4 gene for the analysis of population genetic of two Xiphinema species.  相似文献   

14.
A new species in the genus Ditylenchus, D. stenurus n. sp. collected from western Iran, is described and illustrated herein based on morphological and molecular studies. The new species is characterised by a body length of 772 (663–863) μm, delicate stylet 6 (5–7) μm long, six lines in the lateral field. Median bulb of pharynx well-developed, muscular with crescentic valve. Post-vulval uterine sac well-developed, 35 (30–45) μm long, female tail elongate-conoid, becoming narrow suddenly with finely rounded terminus. The new species comes close in morphology and morphometrics to five known species of the genus, namely D. arachis, D. caudatus, D. clarus, D. myceliophagus, and D. nanus. DNA sequencing data was obtained on the partial 18S, D2/D3 expansion segments of the 28S rRNA gene and internal transcribed spacer (ITS). The phylogenetic relationships of this species with other Ditylenchus spp. using partial 18S–rDNA and D2/D3 indicated that D. stenurus n. sp. clustered together with several species belongs to the D. triformis-group i. e. D. africanus, D. destructor and D. halictus: all sharing a rounded tail terminus and six lines in lateral fields.  相似文献   

15.
Several species of trichodorid nematodes cause economically important disease on plants. Trichodorus comprises most of the trichodorid species. Trichodorus golestanensis n. sp. is described from a forest park in northern Iran. The new species belongs to the T. lusitanicus morpho-species group and is characterized in male by three ventromedian cervical papillae of which two at level of onchiostyle region, three ventromedian precloacal supplements, the posterior one just anterior to retracted spicules and ventrally curved spicules with mid-blade indentation provided with a few bristles. Females are distinguished by well developed triangular vaginal sclerotized pieces in lateral optical view, a rhomboid-shaped vagina and a transverse slit-like vulva in ventral view. Molecular analysis using the sequence of D2-D3 expansion segment of 28S rDNA differentiated the new species and confirmed the relationships of T. golestanensis n. sp. with T. andalusicus and T. asturanus. The sequences of D2-D3 regions were also provided for previous trichodorid records from Iran T. gilanensis, P. teres and N. minor together with discussion of their relationship.  相似文献   

16.
Anguina obesa n. sp., a new species of the genus, causing small seed galls inside the ovaries of foxtail weed plants (Alopecurus mysuroides Huds.) is described and illustrated based on its morphological and molecular characters. The new species is characterized by its 1516–2564 μm long obese females irregularly ventrally curved after fixation, having six lines in lateral fields, 6–9 μm long stylet with well-developed rounded knobs, constriction at junction of isthmus with the pharyngeal bulb, monodelphic-prodelphic female reproductive system, and conical, 60–80 μm long tail. Males of the new species are characterized with their slender 936–1420 μm long body, 25–30 μm long tylenchoid spicules, and bursa not reaching tail tip. Second stage juveniles of the new species were also common inside the galls and also recovered from soil in type locality. The new species is morphologically close to Anguina agropyronifloris, A. amsinckiae, A. paludicola and A. tumefaciens, but is more closely related to A. paludicola, from which it can be separated based on differences in morphological characters and internal transcribed spacer sequence. In Bayesian inference using sequences of the aforementioned genomic fragment, the new species formed a clade with A. agrostis, A. funesta, A. graminis, A. phalaridis and some unidentified isolates, with robust Bayesian posterior probability (BPP). The morphologically closest species, A. paludicola, occupied a separate position, outside of the clade containing the new species. The sequences of two other genomic fragments, 18S and 28S rDNA (D2/D3 region) were also made available for the new species. Morphological comparisons of the new species with the related species are discussed.  相似文献   

17.
Cryphonectriaceae species cause serious canker diseases on chestnut, oak and eucalypt trees. Recently, canker symptoms with typical orange fruiting bodies were observed on Chinese chestnut and oak trees in Hebei, Hubei, Shaanxi and Shandong Provinces in China. In the present study, isolates of these fungi were identified based on phylogenetic and morphological evidence, and their pathogenicities were tested on detached chestnut (Castanea mollissima) branches. DNA sequence comparisons of the internal transcribed spacer (ITS) regions and two regions of β-tubulin (TUB1/TUB2) indicate that these isolates represent five species in the Cryphonectriaceae, viz. Cryphonectria japonica, Cryphonectria parasitica, Aurantiosacculus castaneae sp. nov., Cryphonectria neoparasitica sp. nov. and Endothia chinensis sp. nov. The sexual morph of Aurantiosacculus is discovered for the first time and can be distinguished from the other genera in Cryphonectriaceae by dark brown ascospores and tubiform appendages at both ends. Cryphonectria neoparasitica sp. nov. is different from the other Cryphonectria species by its aseptate ascospores. Endothia chinensis sp. nov. is the sole species in Endothia infecting the host genus Castanea. Additionally, it is much smaller than E. gyrosa and narrower than E. singularis in ascospores. The inoculation results showed that these five Cryphonectriaceae fungi isolated from chestnut or oak are all pathogenic to tested chestnut branches. Cryphonectria parasitica appears to be the most aggressive fungus, followed by C. neoparasitica sp. nov., C. japonica, E. chinensis sp. nov. and A. castaneae sp. nov.  相似文献   

18.
Loquat (Eriobotrya japonica) is an important subtropical fruit crop in Spain and other Mediterranean countries. In recent years, characteristic symptoms of branch canker and dieback have been observed in the main cultivated areas of loquat in Spain. The goal of this study was to identify and characterize the species of Botryosphaeriaceae associated with these symptoms. For this, 36 affected orchards were surveyed between 2010 and 2011 in six provinces of southeastern Spain. Eighty‐four isolates belonging to the family Botryosphaeriaceae were recovered from samples with symptoms. These isolates were characterized by means of phenotypical studies, DNA sequence analyses of the internal transcribed spacer (ITS) and part of the translation elongation factor 1‐α regions, and pathogenicity tests. Ten fungal species were identified: Diplodia malorum, Diplodia olivarum, Diplodia seriata, Diplodia pseudoseriata/Diplodia alatafructa, Diplodia sp., Dothiorella sarmentorum, Neofusicoccum mediterraneum, N. parvum, Spencermartinsia plurivora and S. viticola. In addition, Diplodia eriobotryicola and Dothiorella eriobotryae are newly described. The most frequent species isolated from cankers was D. seriata and, as far as is known, this is the first report of D. malorum, and species belonging to the complex D. pseudoseriata/D. alatafructa, in Spain. All species were pathogenic to 1‐year‐old loquat plants cv. Algerie, with Diplodia sp. and S. viticola as the most virulent.  相似文献   

19.
The following 14 species of phytoseiid mites were recorded on various plants:Typhlodromus kerkirae sp. nov.,T. psyllakisi sp. nov.,T. athenas sp. nov.,T. aff.recki Wainstein,T. cryptus Athias-Henriot,T. atticus sp. nov.,T. longipalpus sp. nov.,T. tiliaruml Oudemans,Seiulus isotrichus Athias-Henriot,Amblyseius aberrans (Oudemans),A. aff.finlandicus (Oudemans),A. stipulatus Athias-Henriot,Phytoseiulus persimilis Athias-Henriot, andPhytoseius (Phytoseius) finitimus Ribaga.  相似文献   

20.
2016年11月,深圳口岸从日本进境罗汉松中截获一种剑线虫。采用贝曼漏斗法分离线虫,结合形态学、28S rRNA基因序列测定以及构建系统发育树,对分离的线虫进行鉴定。该线虫的28SrRNA基因序列和GenBank数据库中的移去剑线虫Xiphinema elongatum(登录号分别为EF140790、KF430803、KF430802和KF430801)的序列同源性高达99%;其形态特征和测计值与X.elongatum最为吻合;系统发育树表明该线虫和X.elongatum聚为一组。通过以上结果判定截获的线虫为移去剑线虫。剑线虫是一种重要的植物外寄生线虫,我国口岸曾经多次截获剑线虫,剑线虫入侵我国的风险较高。本文首次对日本罗汉松中的移去剑线虫进行了详细的描述,提供了该线虫新的地理分布信息。  相似文献   

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